Rapid PCR-based assay for Sclerotinia sclerotiorum detection on soybean seeds

• Main Authors: Edilaine Mauricia Gelinski Grabicoski, David de Souza Jaccoud Filho, Marcos Pileggi, Luciane Henneberg, Marcelo Luiz Cunha Pierre, Cláudio Mauricio Vrisman, Audrei Nisio Gebieluca Dabul
• Format: Article
• Language: English
• Published: Universidade de São Paulo 2015-02-01
• Series: Scientia Agricola
• Subjects: white mold; dissemination; soaking seeds; diagnostic
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-90162015000100069&lng=en&tlng=en

Caused by Sclerotinia sclerotiorum, white mold is an important seed-transmitted disease of soybean (Glycine max). Incubation-based methods available for the detection and quantification of seed-borne inoculum such as the blotter test, paper roll and Neon-S assay are time-consuming, laborious, and not always sensitive. In this study, we developed and evaluated a molecular assay for the detection of S. sclerotiorum in soybean seeds using a species-specific PCR (polymerase chain reaction) primer set and seed soaking (without DNA extraction) for up to 72 h. The PCR products were amplified in all the samples infected with the pathogen, but not in the other samples of plant material or the other seed-borne fungi DNA. The minimum amount of DNA detected was 10 pg, or one artificially infested seed in a 400-seed sample (0.25 % fungal incidence) and one naturally infected seed in a 300-seed sample (0.33 % incidence). The PCR-based assay was rapid (< 9 h), did not require DNA extraction and was very sensitive.